Wavelength-Dependent Damage to Adenoviral Proteins Across the Germicidal UV Spectrum.

Adenovirus, a waterborne pathogen responsible for causing bronchitis, pneumonia, and gastrointestinal infections, is highly resistant to UV disinfection and therefore drives the virus disinfection regulations set by the U.S. Environmental Protection Agency. Polychromatic UV irradiation has been shown to be more effective at inactivating adenovirus and other viruses than traditional monochromatic irradiation emitted at 254 nm; the enhanced efficacy has been attributed to UV-induced damage to viral proteins. This research shows UV-induced damage to adenoviral proteins across the germicidal UV spectrum at wavelength intervals between 200 and 300 nm. A deuterium lamp with bandpass filters and UV light-emitting diodes (UV LEDs) isolated wavelengths in approximate 10 nm intervals. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and image densitometry were used to detect signatures for the hexon, penton, fiber, minor capsid, and core proteins. The greatest loss of protein signature, indicating damage to viral proteins, occurred below 240 nm. Hexon and penton proteins exposed to a dose of 28 mJ/cm2 emitted at 214 nm were approximately 4 times as sensitive and fiber proteins approximately 3 times as sensitive as those exposed to a dose of 50 mJ/cm2 emitted at 254 nm. At 220 nm, a dose of 38 mJ/cm2 reduced the hexon and penton protein quantities to approximately 33% and 31% of the original amounts, respectively. In contrast, a much higher dose of 400 mJ/cm2 emitted at 261 and 278 nm reduced the original protein quantity to between 66-89% and 80-93%, respectively. No significant damage was seen with a dose of 400 mJ/cm2 at 254 nm. This research directly correlates enhanced inactivation at low wavelengths with adenoviral protein damage at those wavelengths, adding fundamental insight into the mechanisms of inactivation of polychromatic germicidal UV irradiation for improving UV water disinfection.

Evaluation of DNA damage reversal during medium-pressure UV disinfection.

Ultraviolet (UV) disinfection relies on the principal that DNA exposure to UV irradiation leads to the formation of cytotoxic lesions resulting in the inactivation of microorganisms. Cyclobutane pyrimdine dimers (CPDs) account for the majority of DNA lesions upon UV exposure. Past research has demonstrated reversal of CPDs in extracted DNA formed at high UV-C wavelength irradiation (280 nm) upon subsequent irradiation at lower UVC wavelengths (230-240 nm). Medium-pressure (MP) UV lamps produce a polychromatic emission giving rise to the possibility that cellular DNA in a target pathogen may undergo simultaneous damage and repair when exposed to multiple wavelengths during the disinfection process, decreasing the efficiency of MP UV lamp disinfection. Culture techniques and a quantitative polymerase chain reaction (qPCR) assay were used to examine cell viability and DNA damage reversal. qPCR results indicated direct photoreversal of UV-induced DNA damage through sequential irradiations of 280 nm followed by 228 nm in Escherichia coli DNA. However, significant photoreversal was only observed after high initial doses and secondary doses of UV light. The doses where significant photoreversal took place were more than 10 times higher than those typically used in UV disinfection. Despite evidence of CPD photoreversal, bacterial growth assays showed no indication that sequential-wavelength irradiations result in higher survival rates than single-wavelength irradiations.